Early transcriptome changes associated with western diet induced NASH in Ldlr−/− mice points to activation of hepatic macrophages and an acute phase response

BackgroundNonalcoholic fatty liver disease (NAFLD) is a global health problem. Identifying early gene indicators contributing to the onset and progression of NAFLD has the potential to develop novel targets for early therapeutic intervention. We report on the early and late transcriptomic signatures of western diet (WD)-induced nonalcoholic steatohepatitis (NASH) in female and male Ldlr−/− mice, with time-points at 1 week and 40 weeks on the WD. Control Ldlr−/− mice were maintained on a low-fat diet (LFD) for 1 and 40 weeks.MethodsThe approach included quantitation of anthropometric and hepatic histology markers of disease as well as the hepatic transcriptome.ResultsOnly mice fed the WD for 40 weeks revealed evidence of NASH, i.e., hepatic steatosis and fibrosis. RNASeq transcriptome analysis, however, revealed multiple cell-specific changes in gene expression after 1 week that persisted to 40 weeks on the WD. These early markers of disease include induction of acute phase response (Saa1-2, Orm2), fibrosis (Col1A1, Col1A2, TGFβ) and NASH associated macrophage (NAM, i.e., Trem2 high, Mmp12 low). We also noted the induction of transcripts associated with metabolic syndrome, including Mmp12, Trem2, Gpnmb, Lgals3 and Lpl. Finally, 1 week of WD feeding was sufficient to significantly induce TNFα, a cytokine involved in both hepatic and systemic inflammation.ConclusionThis study revealed early onset changes in the hepatic transcriptome that develop well before any anthropometric or histological evidence of NALFD or NASH and pointed to cell-specific targeting for the prevention of disease progression

Nrf2 status affects tumor growth, HDAC3 gene promoter associations, and the response to sulforaphane in the colon

BACKGROUND: The dietary agent sulforaphane (SFN) has been reported to induce nuclear factor erythroid 2 (NF-E2)-related factor 2 (Nrf2)-dependent pathways as well as inhibiting histone deacetylase (HDAC) activity. The current investigation sought to examine the relationships between Nrf2 status and HDAC expression in preclinical and translational studies. RESULTS: Wild type (WT) and Nrf2-deficient (Nrf2(−/+)) mice were treated with the colon carcinogen 1,2-dimethylhydrazine (DMH) followed by 400 ppm SFN in the diet (n = 35 mice/group). WT mice were more susceptible than Nrf2(−/+) mice to tumor induction in the colon. Tumors from WT mice had higher HDAC levels globally and locally on genes such as cyclin-dependant kinase inhibitor 2a (Cdkn2a/p16) that were dysregulated during tumor development. The average tumor burden was reduced by SFN from 62.7 to 26.0 mm(3) in WT mice and from 14.6 to 11.7 mm(3) in Nrf2(−/+) mice. The decreased antitumor activity of SFN in Nrf2(−/+) mice coincided with attenuated Cdkn2a promoter interactions involving HDAC3. HDAC3 knockdown in human colon cancer cells recapitulated the effects of SFN on p16 induction. Human subjects given a broccoli sprout extract supplement (200 μmol SFN equivalents), or reporting more than five cruciferous vegetable servings per week, had increased p16 expression that was inversely associated with HDAC3 in circulating peripheral blood mononuclear cells (PBMCs) and in biopsies obtained during screening colonoscopy. CONCLUSIONS: Nrf2 expression varies widely in both normal human colon and human colon cancers and likely contributes to the overall rate of tumor growth in the large intestine. It remains to be determined whether this influences global HDAC protein expression levels, as well as local HDAC interactions on genes dysregulated during human colon tumor development. If corroborated in future studies, Nrf2 status might serve as a biomarker of HDAC inhibitor efficacy in clinical trials using single agent or combination modalities to slow, halt, or regress the progression to later stages of solid tumors and hematological malignancies. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13148-015-0132-y) contains supplementary material, which is available to authorized users

Specific Expression of Anaplasma marginale Major Surface Protein 2 Salivary Gland Variants Occurs in the Midgut and Is an Early Event during Tick Transmission

Infectivity of Anaplasma spp. develops when infected ticks feed on a mammalian host (transmission feed). Specific Anaplasma marginale major surface protein 2 (MSP2) variants are selected for within the tick and are expressed within the salivary glands. The aims of this study were to determine when and where MSP2 variant selection occurs in the tick, how MSP2 expression is regulated in salivary glands of transmission-feeding ticks, and whether the number of A. marginale organisms per salivary gland is significantly increased during transmission feeding. The South Idaho strain of A. marginale was used, as MSP2 expression is restricted to two variants, SGV1 and SGV2, in Dermacentor andersoni. Using Western blot, real-time PCR, and DNA sequencing analyses it was shown that restriction and expression of MSP2 occurs early in the midgut within the first 48 h of the blood meal, when ticks acquire infection. A. marginale is present in the tick salivary glands before transmission feeding is initiated, but the msp2 mRNA and MSP2 protein levels per A. marginale organism increase only minimally and transiently in salivary glands of transmission-feeding ticks compared to that of unfed ticks. A. marginale numbers per tick increase gradually in salivary glands of both transmission-fed and unfed ticks. It is concluded that MSP2 variant selection is an early event in the tick and that MSP2 variants SGV1 and SGV2 are expressed both in the midgut and salivary glands. While MSP2 may be required for infectivity, there is no strict temporal correlation between MSP2 expression and the development of infectivity

Lipidomic and transcriptomic analysis of western diet-induced nonalcoholic steatohepatitis (NASH) in female Ldlr -/- mice.

BackgroundNonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease worldwide, particularly in obese and type 2 diabetic individuals. NAFLD ranges in severity from benign steatosis to nonalcoholic steatohepatitis (NASH); and NASH can progress to cirrhosis, primary hepatocellular carcinoma (HCC) and liver failure. As such, NAFLD has emerged as a major public health concern. Herein, we used a lipidomic and transcriptomic approach to identify lipid markers associated with western diet (WD) induced NASH in female mice.MethodsFemale mice (low-density lipoprotein receptor null (Ldlr -/-) were fed a reference or WD diet for 38 and 46 weeks. Transcriptomic and lipidomic approaches, coupled with statistical analyses, were used to identify associations between major NASH markers and transcriptomic & lipidomic markers.ResultsThe WD induced all major hallmarks of NASH in female Ldlr -/- mice, including steatosis (SFA, MUFA, MUFA-containing di- and triacylglycerols), inflammation (TNFα), oxidative stress (Ncf2), and fibrosis (Col1A). The WD also increased transcripts associated with membrane remodeling (LpCat), apoptosis & autophagy (Casp1, CtsS), hedgehog (Taz) & notch signaling (Hey1), epithelial-mesenchymal transition (S1004A) and cancer (Gpc3). WD feeding, however, suppressed the expression of the hedgehog inhibitory protein (Hhip), and enzymes involved in triglyceride catabolism (Tgh/Ces3, Ces1g), as well as the hepatic abundance of C18-22 PUFA-containing phosphoglycerolipids (GpCho, GpEtn, GpSer, GpIns). WD feeding also increased hepatic cyclooxygenase (Cox1 & 2) expression and pro-inflammatory ω6 PUFA-derived oxylipins (PGE2), as well as lipid markers of oxidative stress (8-iso-PGF2α). The WD suppressed the hepatic abundance of reparative oxylipins (19, 20-DiHDPA) as well as the expression of enzymes involved in fatty epoxide metabolism (Cyp2C, Ephx).ConclusionWD-induced NASH in female Ldlr -/- mice was characterized by a massive increase in hepatic neutral and membrane lipids containing SFA and MUFA and a loss of C18-22 PUFA-containing membrane lipids. Moreover, the WD increased hepatic pro-inflammatory oxylipins and suppressed the hepatic abundance of reparative oxylipins. Such global changes in the type and abundance of hepatic lipids likely contributes to tissue remodeling and NASH severity

Nachrichtendienstpsychologie 3

Der Versuch, nachrichtendienstliche Fragestellungen unter psychologischen Aspekten zu betrachten, ist auf ein positives Interesse gestoßen. In den beiden Bänden Nachrichtendienstpsychologie 1 (Litzcke, 2003) und 2 (Schwan, 2004) der FH Bund erschienen eine Reihe von Aufsätzen, die ein breites Themenspektrum behandeln. Mit Band 1 wurde eine Struktur eingeführt, die sich bewährt hat, und die daher auch für Band 3 beibehalten wird. Die Beiträge des Bandes 3 lassen sich wie folgt zuordnen: 1) Anwendungen vorhandener Konzepte der Psychologie und aus Nachbardisziplinen in den Nachrichtendiensten. Siehe hierzu die Beiträge von Scherer zur Moderation von Qualitätszirkeln im Bundesnachrichtendienst, von Damm und Litzcke zur Akzeptanz des Sicherheitsakteurs „Europa“, von Schwan zur Bedeutung aggressiven Verhaltens für die Arbeit der Nachrichtendienste, von Löhr zum kognitiven Interview und von Wiesen zu den Möglichkeiten einer Gesprächssteuerung. 2) Übertragung psychologischer Methoden auf das Forschungsgebiet Nachrichtendienste zur Gewinnung neuer Ergebnisse / Konzepte. Siehe hierzu die Beiträge von Müller-Enbergs zu den Motiven für eine geheimdienstliche oder nachrichtendienstliche Kooperation bei den inoffiziellen Mitarbeitern der DDR-Staatssicherheit, von Horn zu den Einsatzmöglichkeiten der Operativen Fallanalyse und von Schmalzl zum Konstrukt Einsatzkompetenz. 3) Generierung eigenständiger psychologischer Erkenntnisse und Methoden im Bereich Nachrichtendienste. Dieser Rubrik ist kein Beitrag des Bandes 3 zuzuordnen. Der Abschlussbeitrag von Freitag zur Anwendbarkeit von Täuschungsprinzipien der Zauberkunst in der nachrichtendienstlichen Arbeit entzieht sich dieser Struktur. Der Beitrag ist mit Augenzwinkern geschrieben und soll auch so gelesen werden. Die Themen der Aufsätze sind nicht auf rein nachrichtendienstliche Aufgabenstellungen beschränkt. So werden beispielsweise auch Querschnittsproblematiken behandelt, die nicht nur einem Arbeitsbereich innerhalb der Nachrichtendienste zuzuordnen wären. Damit werden auch Themen behandelt, die ebenso in anderen Behörden und Institutionen von praktischer Relevanz sein können. So ist beispielsweise die Einrichtung von Qualitätszirkeln kein spezifisches Thema der Nachrichtendienste. Da sich aber auch Nachrichtendienste mit der Qualitätssteigerung ihrer Arbeit befassen, ist es angezeigt, entsprechende Ausarbeitungen in die Reihe Nachrichtendienstpsychologie zu integrieren. Auch in Band 3 werden interdisziplinäre Ansätze und Verknüpfungen mit der Polizeipsychologie vorgestellt, auch wenn die dort entwickelten Erkenntnisse und Methoden nicht ungeprüft auf nachrichtendienstliche Aufgabenstellungen übertragen werden können. Gleichwohl können sich aus dem Kontakt zur Polizeipsychologie Anregungen für die Nachrichtendienstpsychologie ergeben

Expression of Anaplasma marginale Major Surface Protein 2 Operon-Associated Proteins during Mammalian and Arthropod Infection

The antigenically variant major surface protein 2 (MSP2) of Anaplasma marginale is expressed from a 3.5-kb operon that contains, in a 5′-to-3′ direction, four open reading frames, opag3 , opag2 , opag1 , and msp2 . This operon structure was shown to be conserved among genotypically and phenotypically distinct A. marginale , A. ovis , and A. centrale strains . The individual OpAG amino acid sequences are highly conserved among A. marginale strains, with identities ranging from 95 to 99%. OpAG2 and OpAG3 were expressed by all examined A. marginale strains during the acute rickettsemia in the mammalian host and, like MSP2, localize to the bacterial surface. OpAG2 and OpAG3 were also expressed in an infected Ixodes scapularis tick cell line. In contrast, the same A. marginale strains expressed only OpAG2 in two different Dermacentor spp. during transmission feeding. OpAG1 expression was not detected in the infected mammalian host, the infected tick cell line, or within infected Dermacentor ticks. The differential expression of outer membrane proteins from within an operon is a novel finding in tick-transmitted bacteria, and the regulation of expression may be broadly applicable to understanding how the pathogen adapts to the mammalian host-tick vector transition

Markers induced or repressed by the western diet, [WD versus LFD] determined by the volcano plot analysis.

Markers induced or repressed by the western diet, [WD versus LFD] determined by the volcano plot analysis.</p

WD effects on hepatic gene expression: Oxidative stress, fibrosis, extracellular matrix remodeling and cell growth.

Hepatic mRNA abundance was quantified by qRTPCR. A and B: Results in the heat maps are presented as the mean value for mRNA abundance of each transcript. The transcripts presented in the heat maps include Cybβ, cytochrome B245 β-chain; Gstα1, glutathione S transferase α1; Hmox1, heme oxygenase 1; Casp1, caspase 1; CtsB, cathepsin B; CtsS, cathepsin S; Col1A1, collagen 1A1; LysOx, lysyl oxidase; Tgfβ1, transforming growth factor β1; Mmp12, matrix metalloprotein 12; Mmp13, matrix metalloprotein 13, Timp1, tissue inhibitor of metalloproteases 1; Bcl2, B-cell lymphoma 2; Gadd45, growth arrest and DNA damage inducible; Gpc3, glypican 3; Hey1, Hes family bHLH transcription factor with YRPW motif1; HeyL, Hes family bHLH transcription factor with YRPW motif 1-like; Ihh, Indian hedgehog; Hhip, hedgehog interacting protein; Snail1, snail family transcriptional repressor; Sox4, SRY-box transcription factor 4. In the heat maps, statistical significance (p-value, pFig 20. C: Ratio of hepatic reduced glutathione (GSH) and oxidized glutathione (GSSG): GSH/GSSG. The results are reported as the mean ± SEM. The “dash” line in the graph indicates that hepatic levels of GSH and GSSG are equal. In the GSH/GSSG graph, statistical significance between values at 4 and 8 wks on the WD versus 1 and 40 wks on the LFD is designated by an asterisk (ӿ). D: the mRNA abundance for cathepsin S (CtsS) is presented as mRNA Abundance Delta Ct, mean ± SEM. Statistical differences in CtsS mRNA abundance after 40 wks on the WD versus LFD is designated by an asterisk (ӿ).</p